|
Status |
Public on Jun 16, 2019 |
Title |
BMDM KO CM1 |
Sample type |
RNA |
|
|
Source name |
BMDM KO CM
|
Organism |
Mus musculus |
Characteristics |
cell type: BMDM genotype: Camkk2 -/- culture condition: CM
|
Treatment protocol |
30% L929-conditioned medium in the presence or absence of 20% of E0771-conditioned medium (CM and RM, respectively)
|
Growth protocol |
BM cells were cultured for 5 days in Corning® Costar® Ultra-Low attachment multi-well plates (Sigma). To establish the purity of BMDM, cells were double-stained with anti-CD11b and anti-F4/80 antibodies. Based on this analysis, more than 95% of cells co-expressed these markers, which is a characteristic of macrophages. In the same experiments, tumor-conditioned medium (20% v/v) was added to differentiation medium.
|
Extracted molecule |
total RNA |
Extraction protocol |
Qiagen Mini Kit Extraction with on-column Dnase treatment followed by Affymatrix WT plus (according to manufacture instructions). Total RNA were assessed for quality with Agilent 2100 Bioanalyzer G2939A (Agilent Technologies,Santa Clara, CA) and Nanodrop 8000 spectrophotometer (Thermo Scientific/Nanodrop, Wilmington, DE). and
|
Label |
Biotin
|
Label protocol |
GeneChip WT Terminal Labeling Kit (Part Number 900670, 900671)
|
|
|
Hybridization protocol |
Hybridization targets were prepared with the Affymetrix WT Plus Kit (Affymetrix, Santa Clara, CA) and Affymetrix GeneChip WT Terminal Labeling Kit (included with Affy WT Plus) from total RNA
|
Scan protocol |
Affymetrix GeneChip® Scanner 7G according to standard Affymetrix GeneChip® Hybridization, Wash, and Stain protocols. (Affymetrix, Santa Clara,CA)
|
Description |
BMDM generated in regular macrophage differentiation medium in the presence of 20% of tumor conditioned medium
|
Data processing |
affymetrix-algorithm-name = sst-rma-gene-full affymetrix-algorithm-version = 1.0 program-name = Expression Console
|
|
|
Submission date |
Oct 30, 2017 |
Last update date |
Jun 16, 2019 |
Contact name |
LUIGI RACIOPPI |
E-mail(s) |
luigi.racioppi@duke.edu
|
Phone |
(919)6816212
|
Organization name |
Duke University
|
Department |
Medicine
|
Lab |
Cell therapy Hematological Malignancies
|
Street address |
La Salle, GSRB1 2006
|
City |
Durham |
State/province |
North Carolina |
ZIP/Postal code |
27707 |
Country |
USA |
|
|
Platform ID |
GPL23038 |
Series (1) |
GSE106357 |
CaMKK2 in myeloid cells is a key regulator of the immune suppressive microenvironment in breast cancer |
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